VEGFC production promoter

ABSTRACT

The purpose of this invention is to find a novel fragrance component effective for prevention or control of swelling, lymphedema, skin aging or obesity from the viewpoint of activation of lymph vessel function by promotion of the expression of VEGFC. This invention provides a swelling improving agent, lymph vessel activator and VEGFC production promoter characterized by containing one or a plurality of components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Divisional of U.S. application Ser. No.15/625,080, filed Jun. 16, 2017, which is a Continuation of U.S.application Ser. No. 14/431,837, which is the U.S. National Stageapplication of PCT/JP2013/076416, filed Sep. 27, 2013, which claimspriority from Japanese application JP 2012-218612, filed Sep. 28, 2012.

TECHNICAL FIELD

The present invention relates to an agent for improving swelling,lymphedema, wrinkles or obesity by promoting activation of lymphaticvessel function by inducing expression of vascular endothelial growthfactor C (VEGFC).

BACKGROUND ART

The cutaneous vasculature is present in the dermis and is composed ofblood vessels and lymph vessels. In order to maintain homeostasis,tissue fluid that has migrated outside blood vessels must berecirculated to veins. Veins present in skin effectively send blood tothe body's core. However, veins per se lack the ability to incorporatetissue fluid. On the basis of this as well, tissue that incorporatestissue fluid, namely lymph vessels, have come to be understood to be anessential structure in the skin as well.

Lymph vessels fulfill an important role in order to maintain themicroenvironment surrounding tissue in a constant state by recoveringunwanted substances present in skin as well as water and proteinconstantly escaping from blood vessels. In addition, it has also come tobe thought to fulfill the role of demonstrating resistance to infectiousfactors and external factors from the outside world by mediating thetransport of T lymphocytes.

Known examples of pathological states associated with lymph vesseldysfunction include symptoms such as swelling and lymphedema (Non-PatentDocument 1: Jusilla, L. & Alitalo, K. (2002), Physiol. Rev. 82,673-700). In addition, lymph vessel function has also been determined tofulfill an important role in not only swelling, but also photoaging(wrinkle formation) of skin accompanying exposure to ultraviolet rays(Non-Patent Document 2: Kajiya, K. & Detmar, M. (2006), J. Invest.Dermatol. 126, 919-21).

According to previous research, VEGFR-3 has been identified as atransmembrane receptor specifically present in lymph vessels, and VEGFCand VEGFD have been found to be ligands thereof. VEGFC activates thefunction of lymph vessels by promoting the proliferation, migration andlumen formation of lymphatic endothelial cells by acting on lymphvessels (Non-Patent Document 1: Jusilla, L. & Alitalo, K. (2002),Physiol. Rev. 82, 673-700). In addition, possibilities are being soughtfor gene therapy involving the application of VEGFC to the pathologicalstate of swelling in the form of edema (Non-Patent Document 3: Saaristo,A., Tammela, T., Timonen, J., Yla-Herttuala, S., Tukianen, E.,Asko-Seljavaara, S. & Alitalo, K. (2004), Faseb. J. 18, 1707-9).

Recently, mutant mice having a genetic mutation that causes lymph vesseldysfunction are known to exhibit obesity as they mature. Findings havebeen obtained regarding the mechanism by which abnormalities in lymphvessel formation and function result in obesity that indicate thatlymphatic fluid flowing through lymph vessels promotes differentiationof mast cell progenitors into fat (Patent Document 4: Harvey, N. L., etal., Nat. Genet. 2005, 37, 1072-81). In other words, functionalabnormalities of lymph vessels have been reported to cause escape oflymphatic fluid outside lymph vessels leading to differentiation intofat and eventually the onset of obesity. Thus, VEGFC promoter isexpected to function as an obesity preventive or therapeutic agent thatfunctionally regenerates lymph vessels.

The VEGF gene family consists of VEGF-A through VEGF-E. Among these,VEGFB and VEGFE have been identified as factors that act only on bloodvessels. Although VEGFA is present in the skin and acts on lymphvessels, it is known to be a factor that conversely exacerbates thefunction of lymph vessels (Non-Patent Document 5: Nagy, J. A., Vasile,E., Feng, D., Sundberg, C., Brown, L. F., Detmar, M. J., Lawitts, J. A.,Benjamin, L., Tan, X., Manseau, E. J., Dvorak, A. M. & Dvorak, H. F.(2002), J. Exp. Med. 196, 1497-506).

In the skin, although VEGFD has been reported to be present in traceamounts in the dermis, since VEGFD knockout mice do not exhibit anyformational or functional abnormalities of skin lymph vesselswhatsoever, VEGFD is not thought to be a factor that is essential forlymph vessel formation in skin (Non-Patent Document 6: Baldwin, M. E.,Halford, M. M., Roufail, S., Williams, R. A., Hibbs, M. L., Grail, D.,Kubo, H., Stacker, S. A. & Achen, M. G. (2005), Mol. Cell. Biol. 25,2441-9). On the other hand, VEGFC in the skin is strongly expressed inthe epidermis. Mice in which VEGFC has been strongly expressed in theepidermis exhibit an increase in the number of lymph vessels present inthe dermis (Non-Patent Document 7: Jeltsch, M., Kaipeinen, A., Joukov,V., Meng, X., Lakso, M., Rauvala, H., Swartz, M., Fukumura, D., Jain, R.K. & Alitalo, K. (1997), Science 276, 1423-5), while on the other hand,when the effects of VEGFC in the epidermis were blocked by highlyexpressing neutralizing antibody of a VEGFC receptor in the form ofVEGFR-3 in the epidermis, findings were obtained that indicated adramatic decrease in the number of lymph vessels in the dermis alongwith the onset of edema (Non-Patent Document 8: Makinen, T., Jussila,L., Veikkola, T., Karpanen, T., Kettunen, M. I., Pulkkanen, K. J.,Kauppinen, R., Jackson, D. G., Kubo, H., Nishikawa, S., Yla-Herttuala,S. & Alitalo, K. (2001), Nat. Med. 7, 199-205). On the basis thereof, ithas been determined that the function of lymph vessels present in skindermis is controlled by VEGFC expressed in the epidermis, and thatswelling (edema) occurs as a result of VEGFC no longer functioning.

It has been previously reported that specific terpenoids and derivativesthereof have a VEGF production-promoting action and are useful as woundhealing agents, hair restorers and hair growth agents and external skinagents having ameliorative effects on skin color (Patent Document 1),and that plant extracts in the manner of apricot kernel extract, ginsengextract, Japanese valerian extract, crataegus fruit extract, melilotextract, white nettle extract and orris root extract have VEGFCproduction-promoting action (Patent Document 2).

PRIOR ART DOCUMENTS Patent Documents

-   [Patent Document 1] Japanese Unexamined Patent Publication No.    2004-2258-   [Patent Document 2] Japanese Unexamined Patent Publication No.    2008-189609

Non-Patent Documents

-   [Non-Patent Document 1] Physiol. Rev. 82, 673-700 (2002)-   [Non-Patent Document 2] J. Invest. Dermatol. 126, 919-21 (2006)-   [Non-Patent Document 3] Faseb. J. 18, 1707-9 (2004)-   [Non-Patent Document 4] Nat. Genet. 37, 1072-81 (2005)-   [Non-Patent Document 5] J. Exp. Med. 196, 1497-506 (2002)-   [Non-Patent Document 6] Mol. Cell. Biol. 25, 2441-9 (2005)-   [Non-Patent Document 7] Science 276, 1423-5 (1997)-   [Non-Patent Document 8] Nat. Med. 7, 199-205 (2001)

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

An object of the present invention is to discover a novel component thatis effective for preventing or controlling swelling, lymphedema, wrinkleformation or obesity by activating lymph vessel function by inducingexpression of VEGFC.

Means for Solving the Problems

As a result of conducting extensive studies towards solving theaforementioned problems, the inventor of the present invention foundthat aromatic components in the manner of rose oxide, citronellylacetate and niaouli oil induce expression of VEGFC. In other words, theinventor of the present invention found that these components thatinduce expression of VEGFC are extremely useful for preventing orinhibiting swelling, lymphedema, formation of wrinkles caused byphotoaging of the skin accompanying exposure to ultraviolet rays, orobesity by promoting activation of lymph vessel function.

The constitution of the present invention is as described below.

[1] A VEGFC production promoter containing one or more componentsselected from the group consisting of rose oxide, citronellyl acetateand niaouli oil.

[2] A lymphangiogenesis and/or lymphatic function promoter containingone or more components selected from the group consisting of rose oxide,citronellyl acetate and niaouli oil.

[3] A swelling ameliorant containing one or more components selectedfrom the group consisting of rose oxide, citronellyl acetate and niaoulioil.

[4] A lymphedema ameliorant containing one or more components selectedfrom the group consisting of rose oxide, citronellyl acetate and niaoulioil.

[5] A wrinkle ameliorant containing one or more components selected fromthe group consisting of rose oxide, citronellyl acetate and niaouli oil.

[6] An obesity ameliorant containing one or more components selectedfrom the group consisting of rose oxide, citronellyl acetate and niaoulioil.

[7] The VEGFC production promoter described in [1], which prevents orinhibits swelling.

[8] The lymphangiogenesis and/or lymphatic function promoter describedin [2], which prevents or inhibits swelling.

[9] The VEGFC production promoter described in [1], which prevents orinhibits lymphedema.

[10] The lymphangiogenesis and/or lymphatic function promoter describedin [2], which prevents or inhibits lymphedema.

[11] The VEGFC production promoter described in [1], which prevents orinhibits wrinkle formation.

[12] The lymphangiogenesis and/or lymphatic function promoter describedin [2], which prevents or inhibits wrinkle formation.

[13] The VEGFC production promoter described in [1], which prevents orinhibits obesity.

[14] The lymphangiogenesis and/or lymphatic function promoter, whichprevents or inhibits obesity.

[15] An aesthetic or therapeutic method for preventing or inhibitingswelling, lymphedema, wrinkles formation and/or obesity, comprisingapplying the VEGFC production promoter described in [1] to the skin of asubject requiring prevention or inhibition of these conditions.

[16] An aesthetic or therapeutic method for preventing or inhibitingswelling, lymphedema, wrinkles formation and/or obesity, comprisingapplying the lymphangiogenesis and/or lymphatic function promoterdescribed in [2] to the skin of a subject requiring prevention orinhibition of these conditions.

[19] A use of the VEGFC production promoter described in [1] forpreventing or inhibiting swelling, lymphedema, wrinkle formation and/orobesity.

[20] A use of the lymphangiogenesis and/or lymphatic function promoterdescribed in [2] for preventing or inhibiting swelling, lymphedema,wrinkle formation and/or obesity.

Effects of the Invention

Expression of vascular endothelial growth factor C (VEGFC) in the bodyis induced and activation of lymph vessel function is promoted byapplying one or more components selected from the group consisting ofrose oxide, citronellyl acetate and niaouli oil. This action makes itpossible to prevent or improve swelling, lymphedema, wrinkle formationor obesity.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the VEGFC-inducing activity of rose oxide,citronellyl acetate and niaouli oil.

MODE FOR CARRYING OUT THE INVENTION Definitions

The term “swelling” in the present description is used in the samecontext as “edema”, and refers to a state observed in subcutaneoustissue in which tissue fluid or lymphatic fluid accumulates in cells,intercellular space or body cavities due to some cause. Moreover, theterm “lymphedema” in the present description refers to a state in whichtissue fluid accumulates in distal regions due to occlusion of lymphvessels.

As is described in detail in the subsequent examples, screening foragents that induce a lymph vessel activating factor in the form of VEGFCwas carried out by allowing various types of candidate aromatics to acton human epidermis-derived cultured cells in the form of HaCaT cells,quantifying VEGFC, and selecting aromatics having VEGFC-inducingactivity as active ingredients by using the amount of VEGFC as anindicator of VEGFC-inducing activity.

As a result, rose oxide, citronellyl acetate and niaouli oil were foundto induce VEGFC.

*Rose Oxide

Rose oxide is an organic compound that has the chemical structureindicated below and has two asymmetric carbons within a moleculethereof.

Rose oxide has a sweet fragrance like that of roses, and is used as apyran-based, monoterpene-based aromatic in cosmetics and soaps. It canbe obtained from roses, rose oil, apricots and peppermint in nature, andhas a plurality of geometrical and optical isomers. Industrially, it isobtained in equal amounts of the cis-form and trans-form by reducingallyl hydroperoxides, which are obtained by photooxidation ofcitronellol, to obtain a diol followed by closing the ring with sulfuricacid. Commercial products available as aromatics consist primarily ofthe cis racemic form.

*Citronellyl Acetate

Citronellyl acetate is an organic compound represented by the followingchemical formula that is in the form of a colorless, clear liquid atnormal temperature.

It is used as an aromatic of foods and cosmetics since it has a uniquefragrance that resembles that of roses or lavender. In nature, it iscontained in essential oils such as citronella oil or geranium oil, andis synthesized industrially from citronellol.

*Niaouli Oil

Niaouli oil is an essential oil collected from the leaves or branches ofMelaleuca virdiflora, a plant of the Myrtaceae family found in thePacific region such as in Indonesia, Australia and New Zealand, and onMadagascar Island in the Indian Ocean. It has bactericidal action and isused in cosmetics, soaps or bath additives and the like.

The aforementioned aromatic components according to the presentinvention are extremely useful for promoting the formation and functionof lymph vessels. Symptoms accompanying lymph vessel dysfunction includenot only swelling and lymphedema, but also photoaging (such as wrinkleformation) of skin accompanying exposure to ultraviolet rays andobesity. The agent according to the present invention is effective forpreventing or inhibiting swelling and lymphedema along with photoagingof the skin accompanying exposure to ultraviolet rays and obesity.Moreover, the aforementioned aromatic components are also thought to beuseful in the treatment of congenital lymphedema.

Photoaging of the skin refers to a change in the appearance and functionof skin typically observed as a result of repeated exposure to sunlight.Constituent elements of sunlight in the form of ultraviolet (UV) rays,and particularly ultraviolet B rays (referred to as UVB rays having awavelength of 290 nm to 320 nm), are the primary cause of photoaging.The exposure level of UVB required to cause photoaging is currently notknown. However, repeated exposure to UVB at a level that causes erythemaor sunburn normally leads to photoaging. In clinical terms, photoagingcan be identified by, for example, rough skin, wrinkle formation,colored spots, pale color, sagging skin, onset of telangiectasia, moleformation, onset of purpura, increased susceptibility to wounds,atrophy, appearance of regions of fibrotic pigment removal and the onsetof premalignant and malignant tumors. Photoaging usually occurs in skinthat has been routinely exposed to sunlight, such as that of the face,ears, head, neck and hands.

The dosage, administration form and dosage form of the VEGFC productionpromoter, lymphangiogenesis and/or lymphatic function promoter, swellingameliorant, lymphedema ameliorant, wrinkle ameliorant or obesityameliorant of the present invention (to also be referred to as the“present agent”) can be suitably determined corresponding to the purposeof use thereof. For example, the aromatic component in the present agentis typically incorporated at 0.0001% by weight to 0.1% by weight,preferably at 0.001% by weight to 0.01% by weight, and optimally at0.005% by weight to 0.01% by weight based on the total weight of thedrug. Although there are no particular limitations on the administrationform of the present agent, and may be administered orally, parenterally,externally or by inhaling and the like, it is preferably a skin externalagent from the viewpoint of being able to act both by inhaling andtranscutaneously. Examples of dosage forms include cosmetics such asperfumes, colognes, shampoos, rinses, skin care products, body shampoos,body rinses, body powders, air fresheners, deodorants, bath additives,lotions, creams, soaps, toothpastes or aerosol products, and other formscommonly used in aromatics. Moreover, the present agent can also be usedin pharmaceuticals such as inhalants.

In addition, the present agent can also suitably incorporate asnecessary other components in addition to the aforementioned essentialcomponents such as components normally used in foods or pharmaceuticalssuch as external skin agents in the manner of skin whitening agents,moisturizers, antioxidants, oily components, ultraviolet absorbers,surfactants, thickeners, alcohols, powdered components, coloring agents,aqueous components, water or various types of skin nutrients.

In addition, other components can also be suitably incorporated,examples of which include sequestering agents such as disodium edetate,trisodium edetate, sodium citrate, sodium polyphosphate, sodiummetaphosphate or gluconic acid, hot water extracts such as caffeine,tannin, verapamil, tranexamic acid and derivatives thereof, licoriceextract, glabridin or quince fruit extract, various types of naturalherbs, drugs such as tocopherol acetate, glycyrrhizic acid andderivatives thereof, and salts thereof, skin whitening agents such asvitamin C, magnesium ascorbyl phosphate, ascorbic acid glucoside,albumin or kojic acid, sugars such as glucose, fructose, mannose,sucrose or trehalose, and vitamin A such as retinoic acid, retinol,retinol acetate or retinol palmitate.

EXAMPLES

Human-derived cultured cells in the form of HaCaT cells (German CancerResearch Center, Heidelberg, Germany) were cultured in DMEM medium(Gibco Low Glucose DMEM™, Life Technologies Inc.) containing 10% FBS(Standard Fetal Bovine Serum, Hyclone Laboratories Inc.) anddisseminated in a 24-well plate to 15×10⁴ cells/well. The medium wasreplaced with serum-free DMEM 24 hours after dissemination followed byculturing for 24 hours. Subsequently, each aromatic dissolved in ethanolwas added (aromatic final concentration: 0.001% to 0.01%). Ethanol wasused as a control. The culture supernatant was recovered after culturingfor 48 hours and the amount of VEGFC in the culture supernatant wasquantified in accordance with the protocol of an ELISA kit (Quanikine®Human VEGF-C Immunoassay (R&D Systems Inc., Minneapolis, Minn.)). Inaddition, alamar blue (TREK Diagnostic Systems, Inc., Cleveland, US) wasadded to the cells after recovering the culture supernatant followed bymeasuring the number of cells based on their fluorescence intensity (Ex:544 nm, Em: 590 nm). The amount of VEGFC in the supernatant was dividedby fluorescence intensity, and the relative VEGFC-inducing activity percell based on a value of 100 for the control (ethanol) is shown inFIG. 1. The synthetic aromatics of rose oxide and citronellyl acetatealong with the natural aromatic, niaouli oil (supplier: Biolandes Inc.),demonstrated VEGFC-inducing activity as shown in FIG. 1 withoutdemonstrating cytotoxicity at each concentration.

FORMULATION EXAMPLES

Although the following indicates formulation examples of the VEGFCproduction promoter of the present invention, carrying out the presentinvention is not limited to the following examples.

Beauty Wash

Formula (wt %) (1) Glycerin 2.0 (2) Dipropylene glycol 2.0 (3) PEG-60hydrogenated castor oil 0.3 (4) Trimethylglycine 0.1 (5) Preservative Assuitable (6) Chelating agent As suitable (7) Dye As suitable (8) VEGFCproduction promoter of present 0.05 invention: Rose oxide (9) Purifiedwater Balance

Beauty Wash

Formula (wt %)  (1) Alcohol 30.0  (2) Butylene glycol 4.0  (3) Glycerin2.0  (4) PPG-13-decyltetradeceth-24 0.3  (5) Octyl methoxycinnamate 0.1 (6) Menthol 0.2  (7) Tocopherol acetate 0.1  (8) Chelating agent Assuitable  (9) Dye As suitable (10) VEGFC production promoter of present0.1 invention: Niaouli oil (11) Purified water Balance

Milky Lotion

Formula (wt %)  (1) Stearic acid 2.0  (2) Cetyl alcohol 1.5  (3)Vaseline 4.0  (4) Squalane 5.0  (5) Glycerol tri-2-ethylhexanoate 2.0 (6) Sorbitan monooleate 2.0  (7) Dipropylene glycol 5.0  (8) PEG15000.3  (9) Triethanolamine 0.1 (10) Preservative As suitable (11) VEGFCproduction promoter of present 0.01 invention: Citronellyl acetate (12)Purified water Balance

Milky Lotion

Formula (wt %)  (1) Ethyl alcohol 10.0  (2) Cyclomethicone 0.1  (3)Butylene glycol 5.0  (4) Dimethicone 3.0  (5) Glycerin 0.1  (6) Menthol1.0  (7) Trimethylsiloxycinnamic acid 0.1  (8) Caffeine 1.0  (9)Trimethylglycine 1.0 (10) Xanthan gum 0.001 (11) Hydroxyethyl cellulose0.1 (12) Fermented soybean extract 1.0 (13) Lauryl betaine 0.5 (14)Carbomer 0.2 (15) Chelating agent As suitable (16) Para-hydroxybenzoateAs suitable (17) Benzoic acid As suitable (18) VEGFC production promoterof present 0.01 invention: Rose oxide (19) Iron oxide As suitable (20)Potassium hydroxide 0.05 (21) Dicalcium glycyrrhizinate 0.01 (22)Pyridoxine hydrochloride 0.01 (23) Ascorbic acid glucoside 0.01 (24)Arbutin 3.0 (25) Saxifraga stolonifera extract 0.1 (26) Water Balance

Milky Lotion

Formula (wt %)  (1) Butylene glycol 4.0  (2) Propylene glycol 4.0  (3)Carbomer 0.2  (4) Potassium hydroxide 0.2  (5) Behenic acid 0.5  (6)Stearic acid 0.5  (7) Isostearic acid 0.5  (8) Glyceryl stearate 1.0 (9) Glyceryl isostearate 1.0 (10) Behenyl alcohol 0.5 (11) Squalane 5.0(12) Trioctanoin 3.0 (13) Phenyl trimethicone 2.0 (14) Batyl alcohol 0.5(15) Dicalcium glycyrrhizinate 0.01 (16) Preservative As suitable (17)Chelating agent As suitable (18) Pigment As suitable (19) VEGFCproduction promoter of present 0.15 invention: Niaouli oil (20) Purifiedwater Balance

Cream

Formula (wt %)  (1) Glycerin 10.0  (2) Butylene glycol 5.0  (3) Carbomer0.1  (4) Potassium hydroxide 0.2  (5) Stearic acid 2.0  (6) Glycerylstearate 2.0  (7) Glyceryl isostearate 2.0  (8) Vaseline 5.0  (9)Preservative As suitable (10) Antioxidant As suitable (11) VEGFCproduction promoter of present 0.02 invention: Citronellyl acetate (12)Purified water Balance (13) Chelating agent As suitable (14) Pigment Assuitable (15) Stearyl alcohol 2.0 (16) Behenyl alcohol 2.0 (17)Hydrogenated palm oil 2.0 (18) Squalane 10.0 (19) Potassium4-methoxysalicylate 3.0

Cream

Formula (wt %)  (1) Glycerin 3.0  (2) Dipropylene glycol 7.0  (3)Polyethylene glycol 3.0  (4) Glyceryl stearate 3.0  (5) Glycerylisostearate 2.0  (6) Stearyl alcohol 2.0  (7) Behenyl alcohol 2.0  (8)Liquid paraffin 7.0  (9) Cyclomethicone 3.0 (10) Dimethicone 1.0 (11)Octyl methoxycinnamate 0.1 (12) Sodium hyaluronate 0.05 (13)Preservative As suitable (14) Antioxidant As suitable (15) VEGFCproduction promoter of present 0.02 invention: Rose oxide (16) Purifiedwater Balance (17) Chelating agent As suitable (18) Pigment As suitable

Gel

Formula (wt %)  (1) Ethyl alcohol 10.0  (2) Glycerin 5.0  (3) Butyleneglycol 5.0  (4) Carbomer 0.5  (5) Aminomethyl propanol 0.3  (6) PEG-60hydrogenated castor oil 0.3  (7) Menthol 0.02  (8) Preservative Assuitable  (9) Chelating agent As suitable (10) VEGFC production promoterof present 0.01 invention: Citronellyl acetate (11) Purified waterBalance

Aerosol

Formula (wt %)  (1) Glycerin 2.0  (2) Dipropylene glycol 2.0  (3) PEG-60hydrogenated castor oil 2.0  (4) HPPCD 1.0  (5) Preservative As suitable (6) Chelating agent As suitable  (7) Dye As suitable  (8) VEGFCproduction promoter of present 0.1 invention: Niaouli oil  (9) Purifiedwater As suitable (10) LPG Balance

Aerosol

Formula (wt %)  (1) Ethanol 60.0  (2) Methyl lactate 0.1  (3) Sodiumlactate 0.1  (4) Tocopherol acetate 0.01  (5) Lactic acid 0.01  (6)Caffeine 0.01  (7) Fennel extract 1.0  (8) Witch hazel extract 1.0  (9)Houttuynia cordata extract 1.0 (10) Dipropylene glycol 1.0 (11) Nitrogengas 0.0 (12) Polyoxyethylene-polyoxypropylene 1.0 decyl tetradecyl ether(13) Butylene glycol 2.0 (14) Tocopherol 0.05 (15) VEGFC productionpromoter of present 0.01 invention (16) PEG-60 hydrogenated castor oil0.1 (17) Water Balance

Fragrance

Formula (wt %) (1) Alcohol 75.0 (2) Purified water Balance (3)Dipropylene glycol 5.0 (4) VEGFC production promoter of present 1.0invention: Citronellyl acetate (5) Antioxidant 8.0 (6) Pigment Assuitable (7) Ultraviolet absorber As suitable

Bath Additive

Formula (wt %) (1) Sodium sulfate 45.0 (2) Sodium bicarbonate 45.0 (3)Lavender oil 9.0 (4) VEGFC production promoter of present 1.0 invention:Niaouli oil

Massage Oil

Formula (wt %)  (1) Erythritol 2.0  (2) Caffeine 5.0  (3) Cork tree barkextract 3.0  (4) Glycerin 50.0  (5) Carboxyvinyl polymer 0.4  (6)Polyethylene glycol 400 30.0  (7) Trisodium edetate 0.1  (8) Polyoxylene(10) methylpolysiloxane 2.0 copolymer  (9) Squalane 1.0 (10) Sodiumhydroxide 0.15 (11) VEGFC production promoter of present 0.01 invention:Rose oxide

Massage Cream

Formula (wt %)  (1) Solid paraffin 5.0  (2) Beeswax 10.0  (3) Vaseline15.0  (4) Liquid paraffin 41.0  (5) 1,3-butyleneglycol 4.0  (6) Glycerinmonostearate 2.0  (7) POE(20) sorbitan monolaurate 2.0  (8) Borax 0.2 (9) Caffeine 2.0 (10) Preservative As suitable (11) Antioxidant Assuitable (12) VEGFC production promoter of present 1.0 invention:Niaouli oil (13) Purified water Balance

The invention claimed is:
 1. A method of ameliorating wrinklescomprising administering to a subject having wrinkles a formulationcomprising one or more components selected from the group consisting ofrose oxide, citronellyl acetate, and niaouli oil, wherein the one ormore components is present in the formulation at a concentration of0.0001 to 0.1% by weight.
 2. The method of claim 1, wherein the one ormore components is present in the formulation at a concentration of0.001 to 0.01% by weight.
 3. The method of claim 1, wherein the one ormore components is present in the formulation at a concentration of0.005 to 0.01% by weight.